Fig. 2

Ameliorated JE progression in NOX2-ablated mice. A, B The frequency and number of Ly-6C ⁺ monocytes and Ly-6G ⁺ neutrophils in the brain. Wild-type (WT) and NOX2 KO mice were inoculated with JEV (7.5 × 10⁷ ffu), and the frequency (A) and total number (B) of Ly-6C ⁺ monocytes and Ly-6G ⁺ neutrophils in the brain were determined by flow cytometric analysis at 3 and 5 dpi using vigorous heart perfusion. Values in the dot-plots show the average percentage of each population after gating on CD11b ⁺ cells. C and D The frequency and number of activated and resting microglia in the brain. The frequency (C) and total number (D) of CD11b⁺CD45^hi (activated microglia) and CD11b⁺CD45^int/lo (resting microglia) cells were determined by flow cytometric analysis at 7th dpi. (E) Total number of lymphoid cells in the brain. The total number of lymphoid cells (CD4 ⁺ and CD8 ⁺ T cells, NK cells) were assessed on the 7th dpi. F Histopathological examinations of the brain tissues derived from infected WT and NOX2 KO mice. Brain sections of WT and NOX2 KO mice were prepared and stained with H&E 5 days after JEV infection via intraperitoneal route. Representative photomicrographs of the brain were obtained from blood vessel areas, meninges, and ventricles. G Inflammatory score of the brain tissues derived from WT and NOX2 KO mice. Inflammation was scored based on the degree of infiltration of inflammatory cells at 5th dpi. H The attenuated expression of inflammatory cytokines and chemokines in NOX2 KO mice. The expression of cytokines and chemokines in the spleen and brain was determined by real-time qRT-PCR at the indicated time points after WT and NOX2 KO mice were inoculated i.p. with JEV (7.5 × 10⁷ ffu). The expression of cytokines and chemokines was normalized to β-actin expression and displayed as the average of at least four independent samples, according to the indicated color on a log₂ scale. Data show the mean ± SEM of the levels derived from at least 2 individual experiments (n = 5–6). *p < 0.05, **p < 0.01, ***p < 0.001 between the levels derived from WT and NOX2 KO mice